Let-7c-5p associate with inhibition of phenobarbital-induced cell proliferation in human palate cells.

Cleft palate (CP) is one of the most common congenital diseases, and is accompanied by a complicated etiology. Medical exposure in women is among one of the reasons leading to CP. Recently, it has been reported that microRNA (miRNA) plays a crucial role in palate formation and the disruption of miRNA that influence the development of CP. Although association with pharmaceuticals and miRNAs were suggested, it has remained largely unknow. The aim of the current investigation is to elucidate upon the miRNA associated with the inhibition of phenobarbital (PB)-induced cell proliferation in human embryonic palatal mesenchymal (HEPM) cells. We showed that PB inhibited HEPM cell viability in a dose-dependent manner. We demonstrated that PB treatment suppressed cyclin-D1 expression in HEPM cells. Furthermore, PB upregulated let-7c-5p expression and downregulated the expression of two downstream genes (BACH1 and PAX3). Finally, we demonstrated that the let-7c-5p inhibitor alleviated PB-induced inhibition of cell proliferation and altered BACH1 and PAX3 expression levels. These results suggest that PB suppresses cell viability by modulating let-7c-5p expression.

Involvement of microRNA-4680-3p against phenytoin-induced cell proliferation inhibition in human palate cells.

Cleft palate (CP) is one of the most common birth defects and is caused by a combination of genetic and/or environmental factors. Environmental factors such as pharmaceutical exposure in pregnant women are known to induce CP. Recently, microRNA (miRNA) was found to be affected by environmental factors. The aim of the present study was to investigate the involvement of miRNA against phenytoin (PHE)-induced inhibition of proliferation in human embryonic palatal mesenchymal (HEPM) cells. We demonstrated that PHE inhibited HEPM cell proliferation in a dose-dependent manner. We found that treatment with PHE downregulated cyclin-D1 and cyclin-E expressions in HEPM cells. Furthermore, PHE increased miR-4680-3p expression and decreased two downstream genes (ERBB2 and JADE1). Importantly, an miR-4680-3p-specific inhibitor restored HEPM cell proliferation and altered expression of ERBB2 and JADE1 in cells treated with PHE. These results suggest that PHE suppresses cell proliferation via modulation of miR-4680-3p expression.

Relationship between phase angle and lower-extremity function in older adults: Itabashi Longitudinal Study on Aging.

OBJECTIVE: Evaluating muscle quality instead of its mass has gained attention in diagnosing sarcopenia. The aim of this study was to examine whether phase angle (PhA) as a bioelectrical impedance analysis (BIA)-derived muscle quality indicator is associated with overall lower extremity function better than appendicular skeletal muscle mass index (ASMI) in community-dwelling older adults. METHODS: This cross-sectional study used data from the Itabashi Longitudinal Study on Aging, a community-based cohort study. A sex-stratified multivariate logistic regression analysis was conducted using PhA and ASMI as exposures, and low physical function defined as short physical performance battery score <10 as the outcome, adjusted for age, being overweight, knee pain, and non-communicable diseases. Discrimination of low physical function was compared using the receiver operating characteristic curve. RESULTS: This study included 1464 participants (age 76 [73-80] y; 757 women), with 58 men (8%) and 66 women (9%) exhibiting low physical function. The multivariate odds ratio (OR; 95% confidence interval [CI]) for low physical function among the highest quartile, compared with the lowest quartile were significant in PhA in multiple sites (e.g., OR, 0.09; 95% CI, 0.03-0.32] for men and 0.12; 95% CI, 0.04-0.33 for women in the left leg) but not in ASMI (OR, 0.51; 95% CI, 0.19-1.34 for men and 0.56; 95% CI, 0.21-1.47 for women). Legs and whole-body PhA outperformed the ASMI in discriminating low physical function (P < 0.001). CONCLUSION: PhA reflected physical function better than ASMI; using PhA instead of ASMI in BIA-based morphometric evaluation may add information on low physical function and enhance the diagnostic value of sarcopenia.

Sasa veitchii extracts protect phenytoin-induced cell proliferation inhibition in human lip mesenchymal cells through modulation of miR-27b-5p.

A cleft lip, with or without a cleft palate, is a common birth defect caused by environmental factors or genetic mutations. Environmental factors, such as pharmaceutical exposure in pregnant women, are known to induce cleft lip, with or without cleft palate in the child. This study aimed to investigate the protective effect of Sasa veitchii extract (SE) on phenytoin-induced inhibition of cell proliferation in human lip mesenchymal cells (KD cells) and human embryonic palatal mesenchymal cells (HEPM cells). We demonstrated that cell proliferation was inhibited by phenytoin in a dose-dependent manner in both KD and HEPM cells. Co-treatment with SE restored phenytoin-induced toxicity in KD cells but did not protect HEPM cells against phenytoin-induced toxicity. Several microRNAs (miR-27b, miR-133b, miR-205, miR-497-5p, and miR-655-3p) is reported to associate with cell proliferation in KD cells. We measured the seven kinds of microRNAs (miR27b-3p, miR-27b-5p, miR-133b, miR-205-3p, miR-205-5p, miR-497-5p, and miR-655-3p) and found that SE suppressed miR-27b-5p induced by phenytoin in KD cells. Furthermore, co-treatment with SE enhanced the expression of miR-27b-5p downstream genes (PAX9, RARA, and SUMO1). These results suggest that SE protects phenytoin-induced cell proliferation inhibition by modulating miR-27b-5p.

Lipomatous mixed tumor of the skin with cystic formation affecting the upper lip: A case report.

Mixed tumor of the skin (MTS) is a rare neoplasm derived from the sweat glands with a reported frequency of 0.01-0.098% among all primary skin tumors. MTS often occurs in the head and neck region and is characterized by a mixture of epithelial, myoepithelial and stromal components. MTS also shows various morphological patterns, thus the presence of variants with rare components and its rarity make the clinical diagnosis even more difficult. A 47-year-old man was referred due to a painless, slowly growing, exophytic swelling intracutaneous mass of the upper lip. Magnetic resonance imaging revealed that the mass was a solid tumor with a fatty component in the proximal portion, while the distal portion was cystic and possibly contained highly viscous fluid. The mass was located between the skin and the orbicularis oris muscle in the upper lip. Excisional biopsy was performed and the lesion showed two intriguing features: A tumor with extensive lipomatous stroma and some large cysts. It was histopathologically diagnosed as lipomatous MTS with cystic formation in the upper lip. No evident signs of recurrence were observed during follow-up. The present report describes this case and includes a brief literature review of reported cases in the lip, since MTS can be confused with various skin lesions in clinical settings due to this rarity. Recognition by clinicians of different variants of MTSs, including the present case, is important for preventing erroneous diagnosis and treatment.

Factors Predicting Tongue Pressure Decline among Community-Dwelling Older Adults: The Takashimadaira Study.

A limited number of longitudinal studies have explored factors contributing to decreases in tongue pressure (TP). This longitudinal study aimed to clarify the factors affecting TP decline among community-dwelling older adults. We followed the Takashimadaira Study participants with a baseline TP ≥ 30 kPa for 2 years. A TP of <30 kPa at follow-up was defined as TP decline. We used Poisson regression with robust standard errors to explore the factors related to TP decline. The studied baseline variables were dental status, sociodemographic characteristics, health behaviors, appetite, medical conditions, physical function, cognitive status, and anthropometric and body composition characteristics. Inverse probability weighting (IPW) was used to adjust for selection bias. Overall, 357 individuals (159 men and 198 women) with a mean (standard deviation) age of 75.9 (4.1) years were included in the analyses. Of these, 59 study participants (16.5%) exhibited TP decline. After adjusting for baseline TP and applying IPW, poor appetite (incident rate ratio [95% confidence interval] = 1.58 [1.01−2.48]), low skeletal muscle mass index (1.66 [1.02−2.70]), and cognitive impairment (1.93 [1.12−3.33]) were associated with TP decline. In conclusion, we demonstrated that baseline appetite, body composition, and cognitive status could predict future TP decline among community-dwelling older adults.

The Semaphorin 3A-AKT axis-mediated cell proliferation in salivary gland morphogenesis and adenoid cystic carcinoma pathogenesis.

We recently demonstrated that Semaphorin 3 A (Sema3A), the expression of which is negatively regulated by Wnt/ß-catenin signaling, promotes odontogenic epithelial cell proliferation, suggesting the involvement of Sema3A in tooth germ development. Salivary glands have a similar developmental process to tooth germ development, in which reciprocal interactions between the oral epithelium and adjacent mesenchyme proceeds via stimulation with several growth factors; however, the role of Sema3A in the development of salivary glands is unknown. There may thus be a common mechanism between epithelial morphogenesis and pathogenesis; however, the role of Sema3A in salivary gland tumors is also unclear. The current study investigated the involvement of Sema3A in submandibular gland (SMG) development and its expression in adenoid cystic carcinoma (ACC) specimens. Quantitative RT-PCR and immunohistochemical analyses revealed that Sema3A was expressed both in epithelium and in mesenchyme in the initial developmental stages of SMG and their expressions were decreased during the developmental processes. Loss-of-function experiments using an inhibitor revealed that Sema3A was required for AKT activation-mediated cellular growth and formation of cleft and bud in SMG rudiment culture. In addition, Wnt/ß-catenin signaling decreased the Sema3A expression in the rudiment culture. ACC arising from salivary glands frequently exhibits malignant potential. Immunohistochemical analyses of tissue specimens obtained from 10 ACC patients showed that Sema3A was hardly observed in non-tumor regions but was strongly expressed in tumor lesions, especially in myoepithelial neoplastic cells, at high frequencies where phosphorylated AKT expression was frequently detected. These results suggest that the Sema3A-AKT axis promotes cell growth, thereby contributing to morphogenesis and pathogenesis, at least in ACC, of salivary glands.

Relationship between Eating Alone and Poor Appetite Using the Simplified Nutritional Appetite Questionnaire.

One prominent factor associated with malnutrition is poor appetite. In Japan, the number of older adults living alone has increased annually. Those living alone tended to eat alone, which may lead to poor appetite. This study aimed to investigate the association between eating alone and poor appetite using an index called the Simplified Nutritional Appetite Questionnaire (SNAQ). We surveyed 818 people aged 70 and over in Takashimadaira, Itabashi-ku, Tokyo, Japan, in 2016. Comparisons were made between two groups, a poor appetite group (n = 295) and a good appetite group (n = 523), and results indicate that the poor appetite group had a higher rate of eating alone than the good appetite group (38.0% vs. 20. 1%: p < 0.001). Multivariable logistic regression (OR; 95%CI) was performed and poor appetite was significantly associated with the Geriatric Depression Scale (GDS) score (1.707; 1.200-2.427), the number of medications (1.061; 1.007-1.118), JST score (0.894; 0.841-0.950), the indication of "very healthy" on a self-rated health scale (0.343; 0.152-0.774), and reports of eating alone (1.751; 1.130-2.712). Our results suggest that eating alone is associated with a poor appetite.

Nutritional status and body composition in cognitively impaired older persons living alone: The Takashimadaira study.

OBJECTIVES: To investigate nutritional status and body composition in cognitively impaired older persons living alone. METHODS: This cross-sectional study included 1051 older adults (633 women and 418 men, mean age: 77.1 years) from the Takashimadaira study. The study participants were categorized according to whether they lived alone, which was confirmed via questionnaire, and had cognitive impairment, which was defined as having a Mini Mental State Examination-Japanese score ≤23. Nutritional status was evaluated using the serum albumin level. The fat-free mass index (FFMI) was calculated based on anthropometric and body composition measurements. A logistic regression model with the outcome of a low serum albumin level (serum albumin <4 g/dL) and low FFMI (<16 kg/m2 in men and <14 kg/m2 in women) were used to analyze the data. RESULTS: The percentages of participants in the living alone (-)/cognitive impairment (-) group, the living alone (+)/cognitive impairment (-) group, the living alone (-)/cognitive impairment (+) group, and the living alone (+)/cognitive impairment (+) group were 54.8%, 37.3%, 5.6%, and 2.3%, respectively. Compared to the living alone (-)/cognitive impairment (-) group, the living alone (+)/cognitive impairment (+) group was more likely to have a low serum albumin level (adjusted odds ratio = 3.10, 95% confidence interval = 1.31 to 7.33) and low FFMI (adjusted odds ratio = 2.79, 95% confidence interval = 1.10 to 7.06) after adjusting for potential confounders. CONCLUSION: Cognitively impaired older adults living alone had poorer nutrition than cognitively normal and cohabitating persons in this study. Our results highlight the importance of paying extra attention to nutritional status for this group of community-dwelling older adults.

MicroRNA-124-3p Plays a Crucial Role in Cleft Palate Induced by Retinoic Acid.

Cleft lip with/without cleft palate (CL/P) is one of the most common congenital birth defects, showing the complexity of both genetic and environmental contributions [e.g., maternal exposure to alcohol, cigarette, and retinoic acid (RA)] in humans. Recent studies suggest that epigenetic factors, including microRNAs (miRs), are altered by various environmental factors. In this study, to investigate whether and how miRs are involved in cleft palate (CP) induced by excessive intake of all-trans RA (atRA), we evaluated top 10 candidate miRs, which were selected through our bioinformatic analyses, in mouse embryonic palatal mesenchymal (MEPM) cells as well as in mouse embryos treated with atRA. Among them, overexpression of miR-27a-3p, miR-27b-3p, and miR-124-3p resulted in the significant reduction of cell proliferation in MEPM cells through the downregulation of CP-associated genes. Notably, we found that excessive atRA upregulated the expression of miR-124-3p, but not of miR-27a-3p and miR-27b-3p, in both in vivo and in vitro. Importantly, treatment with a specific inhibitor for miR-124-3p restored decreased cell proliferation through the normalization of target gene expression in atRA-treated MEPM cells and atRA-exposed mouse embryos, resulting in the rescue of CP in mice. Taken together, our results indicate that atRA causes CP through the induction of miR-124-3p in mice.

Low Dietary Variety and Diabetes Mellitus Are Associated with Frailty among Community-Dwelling Older Japanese Adults: A Cross-Sectional Study.

The association between dietary diversity and frailty remains unknown in older people. We evaluated whether a limited dietary variety is associated with frailty in older adults with diabetes mellitus (DM). This cross-sectional investigation included 1357 adults (median age: 77 years, women: 61.3%). DM was determined by self-reporting, the Dietary Variety Score (DVS) was used to evaluate dietary variety, and the revised Japanese version of the Cardiovascular Health Study criteria evaluated frailty. Participants were divided into 4 groups: no DM/high DVS (non-DMHV), no DM/low DVS (non-DMLV), DM/high DVS (DMHV), and DM/low DVS (DMLV). The prevalence of frailty in each group was 3.6%, 6.7%, 6.7%, and 12.2%. After adjusting for covariates, logistic regression analysis revealed the highest odds ratio (OR) of frailty in the DMLV (non-DMLV, OR = 2.18 (95% confidence interval (CI): 1.25-3.83); DMHV, OR = 1.87 (95% CI: 0.63-5.52); DML, OR = 5.03 (95% CI: 2.05-12.35)). Another logistic regression analysis revealed that a low DVS and DM were independently associated with frailty. Both a low dietary variety and DM were independently related to frailty in older people and the combination increased the prevalence of frailty. These findings suggest that high dietary variety could be important for the prevention of frailty in people with DM.

[Relationship between the rate of a decreased oral function and the nutrient intake in community-dwelling older persons: An examination using oral function-related items in a questionnaire for latter-stage elderly people].

AIM: To determine the rate of a decreased oral function using questions from the Kihon checklist corresponding to the Questionnaire for Latter-stage Elderly People and to clarify nutrient intake in older persons. METHODS: This study targeted 511 older people (217 men, 294 women, average age 73.1±5.6 years old). Their oral function was evaluated using questions on the masticatory function and swallowing function from the Kihon checklist, corresponding to questions on the oral function in the Questionnaire for Latter-stage Elderly People. Participants who had at least one symptom measured were defined as the applicable group (AG). In addition, to evaluate the nutrient intake of the participants, interviews were conducted using the Food Frequency Questionnaire Based on Food Groups. RESULTS: The rate of inclusion in the AG was 32.9% for the total sample, 28.2% for early-stage elderly people, and 40.1% for latter-stage elderly people. The AG rates did not differ significantly between men and women. For latter-stage elderly people, the protein-energy ratio and intakes of total energy, protein, pantothenic acid, folic acid, vitamin B6, niacin, vitamin K, copper, zinc, phosphorus, magnesium, potassium, and total dietary fiber were significantly lower in the AG than in the non-AG. CONCLUSION: The evaluation of placement in the AG through questions on the oral function from the Kihon checklist corresponding to the Questionnaire for Latter-stage Elderly People demonstrated that the rate of a decreased oral function was higher in latter-stage elderly people than in early-stage elderly people. In addition, the latter-stage elderly people in the AG had a lower nutrient intake.

Relationship between Chewing Ability and Nutritional Status in Japanese Older Adults: A Cross-Sectional Study.

OBJECTIVES: This study aimed to determine the relationship between objective chewing ability and the nutritional status of Japanese community-dwelling elders. DESIGN: A cross-sectional study. PARTICIPANTS: A total of 509 community-dwelling elders living in the Tokyo metropolitan area participated in a comprehensive survey conducted in October 2013. MEASUREMENTS: The basic characteristics were sex, age, and body mass index. Undernutrition was examined through serum albumin levels. Chewing ability was examined through color-changeable xylitol gum by evaluating the color changes in chewing gum. Nutritional intake was examined using the semi-quantitative food frequency questionnaire. RESULTS: In the poor chewing ability group, all nutrient intake levels were significantly low, except for carbohydrates, and intake levels for all food groups were significantly low, except for cereals, confectionery, sugars, seasonings, and spices. Additionally, after adjusting for covariates for sex, age, Tokyo Metropolitan Institute of Gerontology-Index of Competence (TMIG-IC) score, Mini-Mental State Examination (MMSE) score, body mass index (BMI), stroke, number of functional teeth, energy intake, and protein intake, chewing ability was found to be significantly associated with undernutrition. CONCLUSION: We concluded that chewing ability was closely associated with nutrient and different food groups' intake, as well as undernutrition, among Japanese community-dwelling elders. Thus, to ensure comprehensive nutritional management, nutritionists and dentists should collaborate when treating the same patients.

Oral frailty and gait performance in community-dwelling older adults: findings from the Takashimadaira study.

Purpose This cross-sectional study compared gait performance between community-dwelling older adults with and without accumulated deficits in oral health, defined as oral frailty.Methods A total of 1,082 individuals (439 men and 643 women; mean age, 77.1 years) from the Takashimadaira study were included in the current analysis. Based on a multifaceted oral health assessment, oral frailty was defined as having three or more of the following six components: (i) fewer teeth, (ii) low masticatory performance, (iii) low articulatory oral motor skills, (iv) low tongue pressure, (v) difficulties in eating, and (vi) swallowing. Eight gait parameters were assessed using an electronic walkway. Gait characteristics comparison between groups with and without oral frailty was performed using multiple linear regression models. Models were adjusted for age, sex, educational status, income, smoking, drinking, physical activity level, height, body mass index, comorbidities, and the presence of chronic pain.Results Oral frailty was observed in 227 (21.0%) participants. After adjusting for potential confounders, the participants with oral frailty had slower gait speed, shorter stride and step length, wider step width, and longer double support duration as well as higher variability of stride length and step length.Conclusions Oral frailty was associated with poor gait performance among community-dwelling older adults.

The Mini Nutritional Assessment-Short Form as a predictor of nursing home mortality in Japan: A 30-month longitudinal study.

OBJECTIVES: We examined whether the Mini Nutritional Assessment-Short Form (MNA®-SF) predicted mortality in 367 nursing home residents (82% women; mean age = 84.4 ± 8.5 years) in Japan. MEASUREMENTS: We examined participants' basic characteristics (sex, age, height, weight, and medical history), the Barthel index (BI), clinical dementia rating (CDR), and six items of the MNA®-SF. The association between the MNA®-SF and 30-month mortality was assessed using a Cox proportional regression analysis. RESULTS: During the study, 157 (42.8%) participants died. MNA®-SF scores in the Survival group were significantly higher than in the Death group (9.4 ± 2.1 vs. 8.4 ± 2.3, respectively; p < .001). After adjusting for age, sex, history of aspiration pneumonia, BI, and CDR, MNA®-SF scores were significantly associated with 30-month mortality (hazard ratio: 0.89, 95% confidence interval: 0.82-0.97, p = .005). CONCLUSION: The MNA®-SF was an effective predictor of mortality among nursing home residents in Japan, even after adjusting for potential confounders. These results indicate that periodically evaluating nutritional status using the MNA®-SF, and nutritional interventions according to status, may result in maintenance and improvement of nutritional status, as well as lead to reduced mortality.

Association between decrease in frequency of going out and oral function in older adults living in major urban areas.

AIM: To examine the association between a decrease in the frequency of going out and oral function in independent older adults living in the urban area of Tokyo. METHODS: The participants analyzed were 785 older adults from the "Takashimadaira Study" (344 men and 441 women, age 77.0 ± 4.6 years). This study investigated the following items: decrease in frequency of going out; basic characteristics (sex, age); physical factors, such as oral function (difficulty chewing, difficulty swallowing, dry mouth); body pain; the Japan Science and Technology Agency Index of Competence; physical activities; psychological factors, such as the Geriatric Depression Scale-15 score; and social and environmental factors, such as the presence or absence of participation in organization activities. RESULTS: To investigate the factors associated with a decrease in frequency of going out, logistic regression analysis showed an association with age (OR 1.08, 95% CI 1.03-1.13), difficulty chewing (OR 2.41, 95% CI 1.52-3.83), dry mouth (OR 1.68, 95% CI 1.07-2.64), body pain (OR 1.78, 95% CI 1.14-2.78), Japan Science and Technology Agency Index of Competence scores (OR 0.91, 95% CI 0.84-0.99), physical activities (OR 0.99, 95% CI 0.98-1.00), Geriatric Depression Scale-15 scores (OR 1.13, 95% CI 1.05-1.21) and organization activities (OR 1.94, 95% CI 1.22-3.07). Covariance structural analyses showed that both "difficulty chewing" and "dry mouth" significantly affected "decrease in frequency of going out." In addition, decrease in frequency of going out was significantly affected by " Geriatric Depression Scale-15 scores" through oral function. CONCLUSIONS: The relationship between oral function and decrease in frequency of going out was clarified, after the multifaceted factors were adjusted. Geriatr Gerontol Int 2019; 19: 792-797.

Relationship between mortality and Council of Nutrition Appetite Questionnaire scores in Japanese nursing home residents.

OBJECTIVE: This 1-y cohort study examined whether Council of Nutrition Appetite Questionnaire (CNAQ) scores predicted mortality in 316 elderly Japanese residents of five nursing homes (60 men, 256 women; mean age: 84.9 ± 8.3 y). METHODS: The baseline survey included participant characteristics (e.g., age, sex, height, weight, and medical history), and Barthel Index (BI), Clinical Dementia Rating (CDR), Mini Nutritional Assessment-Short Form (MNA®-SF), CNAQ, Simplified Nutritional Appetite Questionnaire (SNAQ; simplified CNAQ), and SNAQ for the Japanese elderly (SNAQ-JE) scores. RESULTS: Following the baseline survey, mortality data were collected for 1 y; during this time, 62 participants (19.6%) died. The deceased group's CNAQ scores (25.1 ± 4.8) were significantly lower than those of the survival group (28 ± 3.6; P < 0.001). After adjusting for age, sex, medical history, BI, CDR, and MNA®-SF scores in Cox proportional regression, CNAQ (hazard ratio [HR], 0.91; 95% confidence interval [CI], 0.85-0.97; P = 0.004), SNAQ (HR, 0.84; 95% CI, 0.75-0.93; P = 0.001), and SNAQ-JE (HR, 0.84; 95% CI, 0.76-0.92; P < 0.001) scores were related to mortality. CONCLUSIONS: This study showed that CNAQ scores were inversely associated with 1-y mortality. Furthermore, appetite assessment using the CNAQ predicted the death of Japanese nursing home residents. Similarly, the SNAQ and SNAQ-JE scores were inversely associated with 1-y mortality.

Low-grade myofibroblastic sarcoma arising in the tip of the tongue with intravascular invasion: A case report.

Low-grade myofibroblastic sarcoma (LGMS) is a rare intermediate tumor, which rarely metastasizes and has myofibroblastic differentiation in various sites. It is particularly associated with the tongue in the head and neck region. The lack of any pathological features means it is difficult to make a conclusive diagnosis of LGMS. The immunohistochemical features and genomic rearrangements, including SS18-SSXs and MYH9-USP6s and the genetic mutations of cancer-associated genes, including APC, CTNNB1, EGFR, KRAS, PIK3CA and p53 were examined in a case of LGMS arising in the tip of the tongue. Immunohistochemically, the tumor cells were positive for alpha-smooth muscle actin and vimentin, as in previous reports. They demonstrated neither genomic rearrangements nor point mutations of cancer-associated genes. Although several tumor cells demonstrated intravascular invasion, the MIB-l labeling index of the cells was the same as the original lesion. To the best of our knowledge, this is the first case report of LGMS arising in the tip of the tongue with intravascular invasion.

GLI-mediated Keratin 17 expression promotes tumor cell growth through the anti-apoptotic function in oral squamous cell carcinomas.

PURPOSE: Keratin 17 (KRT17) has been suggested as a potential diagnostic marker of squamous cell carcinoma including oral squamous cell carcinoma (OSCC). The current study was conducted to clarify the function of KRT17 and its expression mechanism in OSCC. METHODS: Immunohistochemical analyses were carried out to examine the expression of KRT17, GLI family zinc finger (GLI)-1, GLI-2, or cleaved caspase-3 in OSCCs. The expression of KRT17, GLI-1, or GLI-2 was investigated among OSCC cell lines, and the effects of loss-of-function of KRT17 or GLI, using siRNA or inhibitor, on the cell growth of the OSCC cell line HSC-2 particularly with respect to apoptosis were examined. RESULTS: Immunohistochemical analyses of tissue specimens obtained from 78 OSCC patients revealed that KRT17 was not observed in non-tumor regions but was strongly expressed at high frequencies in tumor regions. Knockdown of KRT17 increased the number of cleaved caspase-3-positive cells, leading to the reduction of cell number. Loss-of-function of GLI-1 or GLI-2 also increased the cell numbers of apoptotic cells positive for staining of Annexin-V and propidium iodide (PI) and the terminal deoxynucleotidyl transferase dUTP-biotin nick-end labeling (TUNEL) method, and induced DNA fragmentation. This inhibitory effect on cell growth was partially rescued by exogenous KRT17 expression. In the KRT17-positive regions in OSCCs, GLI-1 or GLI-2 was frequently detected, and the number of cells with cleaved caspase-3 positive was decreased. CONCLUSIONS: KRT17 promotes tumor cell growth, at least partially, through its anti-apoptotic effect as a result of the KRT17 overexpression by GLIs in OSCC.

Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) expression and possible function in mouse tooth germ development.

Abnormal expression of Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) is involved in the pathogenesis of FSHD. FRG1 is also important for the normal muscular and vascular development. Our previous study showed that FRG1 is one of the highly expressed genes in the mandible on embryonic day 10.5 (E10.5) than on E12.0. In this study, we investigated the temporospatial expression pattern of FRG1 mRNA and protein during the development of the mouse lower first molar, and also evaluated the subcellular localization of the FRG1 protein in mouse dental epithelial (mDE6) cells. The FRG1 expression was identified in the dental epithelial and mesenchymal cells at the initiation and bud stages. It was detected in the inner enamel epithelium at the cap and early bell stages. At the late bell and root formation stages, these signals were detected in ameloblasts and odontoblasts during the formation of enamel and dentin matrices, respectively. The FRG1 protein was localized in the cytoplasm in the mouse tooth germ in vivo, while FRG1 was detected predominantly in the nucleus and faintly in the cytoplasm in mDE6 cells in vitro. In mDE6 cells treated with bone morphogenetic protein 4 (BMP4), the protein expression of FRG1 increased in cytoplasm, suggesting that FRG1 may translocate to the cytoplasm. These findings suggest that FRG1 is involved in the morphogenesis of the tooth germ, as well as in the formation of enamel and dentin matrices and that FRG1 may play a role in the odontogenesis in the mouse following BMP4 stimulation.